ObjectiveTo systematically compare the differential regulation of the maternal-fetal interface cell lineages and communication networks in the CBA/J×DBA/2 mouse model of recurrent pregnancy loss （RPL） by the two classic therapeutic methods-tonifying the kidney to stabilize the fetus and invigorating the spleen to stabilize the fetus （Shoutai Wan， Juyuan Jian）-of traditional Chinese medicine （TCM） at the single-cell resolution and clarify their modern scientific connotations. MethodsFemale non-pregnant CBA/J mice were caged with male BALB/c （blank group） and DBA/2 （modeling group） mice separately. Pregnant mice in the modeling group were randomly grouped as follows： high/low-dose Shoutai Wan， high/low-dose Juyuan Jian， model （RPL）， and positive control （dydrogesterone）， with 10 mice in each group. Starting from the day after the detection of the vaginal plug， mice were administrated with drugs or an equal volume of normal saline by gavage for 10 consecutive days. After the intervention， the following indicators were measured. ① Macroscopic evaluation： general conditions， uterine wet weight， embryo loss rate， four coagulation parameters ［prothrombin time （PT）， activated partial thromboplastin time （APTT）， fibrinogen （FIB）， and thrombin time （TT）］， and peripheral blood estradiol （E₂） and progesterone （Pg） levels. The decidua with embryos was stained with hematoxylin-eosin （HE） and evaluated by transmission electron microscopy （TEM）. The expression of B-cell lymphoma-2 （Bcl-2）， vascular endothelial growth factor （VEGF）， angiotensin Ⅱ （AngⅡ）， matrix metalloproteinase-2 （MMP-2）， interleukin-6 （IL-6）， leukemia inhibitory factor （LIF）， CXC chemokine ligand 12 （CXCL12）， and microtubule-associated protein 1 light chain 3 homolog （LC3）Ⅰ/Ⅱ was quantified by Western blot. ② Mechanism analysis at the single-cell level： The decidua with embryos from the blank， model， high-dose Shoutai Wan， and high-dose Juyuan Jian groups （6 mice per group， with 3 single-cell samples per group， totaling 24 mice） were analyzed by the BD Rhapsody™ platform， and the whole-cell atlas was drawn by uniform manifold approximation and projection （UMAP） dimensionality reduction clustering combined with the single-cell mouse cell atlas （scMCA）. The differentially expressed genes （DEGs） and cell interaction networks were analyzed via Gene Ontology （GO）， Kyoto Encyclopedia of Genes and Genomes （KEGG）， and CellChat， and the protein-protein interaction （PPI） map of subtype cells was constructed. The CytoTRACE pseudo-temporal analysis was performed to explore the developmental trajectories of core immune cells （natural killer cells， NK cells） from maternal and fetal sources. Results① Pathological and Western blot results indicated that compared with the blank group， the RPL group showed an increase in the embryo loss rate （P<0.01）， down-regulated expression of Bcl-2， LIF， MMP-2， and Vegf in the decidua with embryos （P<0.05）， up-regulated protein levels of CXCL-12， AngⅡ， and IL-6 （P<0.05）， blocked angiogenesis， apoptosis-inflammation imbalance， and coagulation dysfunction. Both prescriptions dose-dependently reduced the abortion rate and restored the angiogenesis-inflammation balance， and Shoutai pill showed superior performance in restoring the E₂ level to the Pg level （P<0.05）. ② Single-cell transcriptome analysis indicated that compared with the blank group， the RPL group showed differences in multiple key cell populations such as decidual cells， trophoblast cells， endothelial cells， erythroblasts， NK cells， and macrophages at the maternal-fetal interface. Immunity and angiogenesis were the key links in RPL. Compared with the RPL group， high-dose Shoutai Wan reversed the changes of NK cells in the embryonic layer （upregulating the mRNA levels of 17 genes and downregulating the mRNA levels of 29 genes） and macrophages （upregulating the mRNA levels of 117 genes and downregulating the mRNA levels of 53 genes） through the regulation of gene expression. High-dose Shoutai pill regulated the immune cells to affect unfolded proteins， cell adhesion， and programmed cell death， thereby promoting decidualization and angiogenesis and modulating embryo-membrane development. High-dose Juyuan Jian regulated the key subgroups of NK cells （up-regulating the mRNA levels of 9 genes and down-regulating the mRNA levels of 17 genes） and macrophages （up-regulating the mRNA levels of 110 genes and down-regulating the mRNA levels of 81 genes）， which affected decidual inflammation and apoptosis and intervened in glycolysis. ③ The pseudo-temporal analysis and communication network indicated that the communication frequency of the RPL group decreased. High-dose Shoutai Wan restored maternal-fetal tolerance through pathways such as NKG2D， CDH5， GDF， and FASLG. High-dose Juyuan Jian enhanced the IL-6/LIFR/JAK/signal transducer and activator of transcription 3 （STAT3） and desmosome/SEMA6/tumor necrosis factor-like weak inducer of apoptosis （TWEAK） signaling to improve endometrial receptivity. The RPL group showed an increased proportion of toxic dNK7， a decreased proportion of reparative dNK4， and blocked embryo fNK1. High-dose Shoutai Wan down-regulated dNK7 and up-regulated dNK4. High-dose Juyuan Jian inhibited the terminal differentiation of dNK7 and up-regulated LILRB1， thus restoring the balance of cytotoxicity and repair. ConclusionBoth the kidney-tonifying and spleen-invigorating methods are effective in treating RPL. NK and macrophages are the key immune cells in the interaction between the embryo and the membrane. The kidney-tonifying method （Shoutai Wan） has an advantage in regulating the phenotypes of unfolded protein， cell adhesion， and programmed cell death， and shows expression characteristics closer to the physiological state in the regulation of NKG2D and CDH5 signals. The spleen-invigorating method （Juyuan Jian） has an advantage in regulating epithelial-mesenchymal transition （EMT）， angiogenesis， and glycolysis and shows higher communication intensity in the IL-6 and LIFR pathways.

ObjectiveTo investigate whether Bushen Huoxue prescription improves embryo implantation through regulating exosomal miRNA to enhance maternal-fetal interface communication based on Bushen Huoxue therapy. MethodsIn the animal experiment， all the rats （except for the blank group） were administered hydroxyurea （450 mg·kg^-1） via gavage for 10 d， as well as epinephrine （0.3 mg·kg^-1） and mifepristone （5.5 mg·kg^-1） via subcutaneous injection for 7 d to establish an implantation disorder model of kidney deficiency and blood stasis type. The Bushen Huoxue prescription （BSHX） groups were administered the prescription at different doses （7.30 g·kg^-1 for the high-dose group， 3.65 g·kg^-1 for the medium-dose group， and 1.83 g·kg^-1 for the low-dose group） via gavage. The dydrogesterone group was administered the corresponding medicine （2.63 mg·kg^-1） via gavage. After intervention for 10 days， uterine histopathological changes were observed via hematoxylin-eosin （HE） staining. Mucin （MUC1）， forkhead box protein O1 （FoxO1）， and homeobox A10 （HoxA10） expression levels were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot. Cell experiment selected primary endometrial epithelial cells （EEC） and trophoblast cells （TC） as research subjects. Exosome-free medicated serum was prepared by ultracentrifugation and cultured in complete medium. Exosomes were isolated from cell supernatants by ultracentrifugation for cross-co-culture. After 48 h， migration and invasion abilities were assessed by scratch and Transwell assays. Sequencing was then performed on EEC-origin exosomal miRNA. ResultsThe model rats exhibited thin endometrium， along with reduced blood vessels， glandules， and pinopode numbers. BSHX improved endometrial morphology and increased pinopode numbers. MUC1， FoxO1， and HoxA10 expressions were downregulated in the model rats， while these parameters were upregulated after BSHX medium- and high-dose intervention. In the cell experiment， after exosome-free medicated serum intervention for 24 h， migration and invasion abilities were enhanced in the BSHX groups （P<0.01）. In EEC-origin exosomal miRNA sequencing， gene ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） analyses revealed enrichment in biological processes （gastrulation， neuronal differentiation， alongside cell development and regeneration）， involving the mitogen-activated protein kinase （MAPK）， FoxO1， Wnt， mammalian target of rapamycin （mTOR）， and tumor necrosis factor （TNF） signaling pathways. ConclusionBSHX promotes embryo implantation by improving endometrial receptivity via regulating exosomal miRNA. These findings provide potential targets for exosomal miRNA-based assisted reproductive strategies and a novel theoretical basis for infertility treatment by traditional Chinese medicine.

To perform the phylogenetic characterization of an isolated porcine rotavirus(PoRV)and investigate its pathogenicity in suckling mice and piglets.A G4P[23]genotype PoRV strain JSJR2023 was successfully isolated from the diarrheic piglet feces through propagation in MA104 cells.The viral proliferation kinetics were analyzed using TCID50 assays,followed by complete genome sequencing through Sanger sequencing platforms.Comprehensive genotyping and phylogenetic reconstruction were conducted using MEGA7.0 with maximum likelihood algorithms.Pathogenicity was assessed in the following animal models:5-day-old C57BL/6 mice and 3-day-old piglets.Multidimensional evaluation included clinical monitoring(diarrhea scoring,growth parameters),virological detection,and histopathological analysis of intestinal tissues.The virus strain JSJR2023 could replicate efficiently in MA104 cells,achieving peak titers of 107.5 TCID50/mL.Whole genome genotype analysis showed that the strain belonged to G4-P[23]-I5-R1-C1-M1-A8-N1-T1-E1-H1.Phylogenetic analysis indicated that the VP3 and NSP4 genes of JSJR2023 strain were most closedrelated to human species rotaviruses,suggesting genetic reassortment between human and porcine RV strains.The animal experiments in suckling mice showed that the JSJR2023 strain infection caused diarrhea symptoms,intestinal edema and congestion,and shedding of intestinal villus epithelial cells.The pathogenicity experiments in piglets showed that compared with the control group,the challenged group of pig-lets had severe diarrhea symptoms,accompanied by reduced appetite and listlessness.Post-mortem examination revealed that the intes-tines were significantly thinner,congested,and filled with yellow watery contents.The challenged piglets showed typical pathological changes such as thinning of the intestinal wall and shortening and shedding of intestinal villi.In conclusion,this study successfully iso-lated a human-porcine recombinant G4P[23]PoRV strain and established the infection models in suckling mice and piglets,providing important tools for investigating the pathogenic mechanism of PoRV,evaluating vaccines and developing antiviral drug.

To perform the phylogenetic characterization of an isolated porcine rotavirus(PoRV)and investigate its pathogenicity in suckling mice and piglets.A G4P[23]genotype PoRV strain JSJR2023 was successfully isolated from the diarrheic piglet feces through propagation in MA104 cells.The viral proliferation kinetics were analyzed using TCID50 assays,followed by complete genome sequencing through Sanger sequencing platforms.Comprehensive genotyping and phylogenetic reconstruction were conducted using MEGA7.0 with maximum likelihood algorithms.Pathogenicity was assessed in the following animal models:5-day-old C57BL/6 mice and 3-day-old piglets.Multidimensional evaluation included clinical monitoring(diarrhea scoring,growth parameters),virological detection,and histopathological analysis of intestinal tissues.The virus strain JSJR2023 could replicate efficiently in MA104 cells,achieving peak titers of 107.5 TCID50/mL.Whole genome genotype analysis showed that the strain belonged to G4-P[23]-I5-R1-C1-M1-A8-N1-T1-E1-H1.Phylogenetic analysis indicated that the VP3 and NSP4 genes of JSJR2023 strain were most closedrelated to human species rotaviruses,suggesting genetic reassortment between human and porcine RV strains.The animal experiments in suckling mice showed that the JSJR2023 strain infection caused diarrhea symptoms,intestinal edema and congestion,and shedding of intestinal villus epithelial cells.The pathogenicity experiments in piglets showed that compared with the control group,the challenged group of pig-lets had severe diarrhea symptoms,accompanied by reduced appetite and listlessness.Post-mortem examination revealed that the intes-tines were significantly thinner,congested,and filled with yellow watery contents.The challenged piglets showed typical pathological changes such as thinning of the intestinal wall and shortening and shedding of intestinal villi.In conclusion,this study successfully iso-lated a human-porcine recombinant G4P[23]PoRV strain and established the infection models in suckling mice and piglets,providing important tools for investigating the pathogenic mechanism of PoRV,evaluating vaccines and developing antiviral drug.

ObjectiveTo explore the mechanism of Bushen Huoxue enema in treating the rat model of kidney deficiency and blood stasis-thin endometrium （KDBS-TE） by transcriptome sequencing. MethodThe rat model of KDBS-TE was established by administration of tripterygium polyglycosides tablets combined with subcutaneous injection of adrenaline. The pathological changes of rat endometrium in each group were then observed. Three uterine tissue specimens from each of the blank group， model group， and Bushen Huoxue enema group were randomly selected for transcriptome sequencing. The differentially expressed circRNAs， lncRNAs， and miRNAs were screened， and the disease-related specific competitive endogenous RNA （ceRNA） regulatory network was constructed. Furthermore， the gene ontology （GO） functional annotation and the Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment were performed for the mRNAs in the network. ResultCompared with the blank group， the model group showed endometrial dysplasia， decreased endometrial thickness and endometrial/total uterine wall thickness ratio （P<0.01）， and differential expression of 18 circRNAs， 410 lncRNAs， and 7 miRNAs. Compared with the model group， the enema and estradiol valerate groups showed improved endometrial morphology and increased endometrial thickness and ratio of endometrial to total uterine wall thickness （P<0.05）. In addition， 21 circRNAs， 518 lncRNAs， and 17 miRNAs were differentially expressed in the enema group. The disease-related specific circRNA-miRNA-mRNA regulatory network composed of 629 nodes and 664 edges contained 2 circRNAs， 34 miRNAs， and 593 mRNAs. The lncRNA-miRNA-mRNA regulatory network composed of 180 nodes and 212 edges contained 5 lncRNAs， 10 miRNAs， and 164 mRNAs. The mNRAs were mainly enriched in Hippo signaling pathway， autophagy-animal， axon guidance， etc. ConclusionBushen Huoxue enema can treat KDBS-TE in rats by regulating specific circRNAs， lncRNAs， and miRNAs in the uterus and the ceRNA network.

Objective:To identify factors influencing the recurrence of psoriasis, and to explore the association between the recurrence of psoriasis and metabolism-related markers.Methods:A retrospective investigation was conducted on the recurrence status of patients with psoriasis vulgaris, who visited the Department of Dermatology, Peking University Third Hospital from January 2016 to April 2023. Patients with recurrence intervals > 3 months were included in the non-rapid recurrence group (non-persistent psoriasis group), while patients with recurrence intervals ≤ 3 months were included in the rapid recurrence group (persistent psoriasis group). General conditions and relapse triggers were analyzed between the two groups. Metabolism-related laboratory data, as well as detection results of serum fatty acid-binding protein 4 (FABP4) and FABP5 in some patients, were collected, and relationships between these indicators and psoriasis recurrence were analyzed. Comparisons between groups were performed using t test, non-parametric test or chi-square test; linear regression analysis was performed to identify possible factors influencing the FABP levels, and multivariate logistic regression analysis to identify relapse triggers. Results:A total of 255 patients were collected, including 194 with non-persistent psoriasis and 61 with persistent psoriasis. There were no significant differences in gender, age (stratified every 30 years), course of psoriasis (stratified every 10 years), family history of psoriasis, and main therapies between the two groups (all P > 0.05). The patients in the non-persistent psoriasis group were more prone to recurrence due to seasonal effects ( χ2 = 18.98, P < 0.001). The proportion of patients with dyslipidemia was significantly higher in the persistent psoriasis group than in the non-persistent psoriasis group ( χ2 = 54.44, P < 0.001). Compared with the non-persistent psoriasis group, the persistent psoriasis group showed significantly increased body mass index and levels of triglycerides, uric acid, and C-reactive protein (all P < 0.05), but significantly decreased high-density lipoprotein cholesterol levels ( U = 3 348.00, P < 0.001). The levels of FABP4 and FABP5 were significantly higher in the persistent psoriasis group than in the non-persistent psoriasis group (both P < 0.05). In the linear regression model adjusted for body mass index and dyslipidemia, FABP4 levels were associated with recurrence status of psoriasis ( P < 0.001). Multivariate logistic regression analysis revealed a significant correlation between dyslipidemia and persistent psoriasis ( P < 0.001) . Conclusion:The psoriasis patients with recurrence intervals ≤ 3 months may be more prone to develop metabolic diseases such as dyslipidemia, and dyslipidemia and elevated FABP4 levels may be associated with the recurrence of psoriasis.

Objective:To systematically evaluate the efficacy of hyperbaric oxygen (HBO) combined with comentional drugs in the treatment of angina pectoris caused by coronary heart disease with meta-analysis.Methods:The data on clinical randomized controlled trials (RCTs) of HBO combined with conventional drugs in the treatment of angina pectoris due to coronary heart disease in Cochrane Library, Medline, Embase, Wanfang Data, Cqvip, China National Knowledge Infrastructure (CNKI) were collected, and the time period was from the respective founding day of the above databases to October 2020. Three researchers independently screened the collected RCTs, extracted relevant data, and evaluated the quality of the trials. RevMan 5.4 software was used for meta-analysis.Results:A total of 11 RCTs were included in this study. Meta-analysis results showed that the HBO group was better than the control group in terms of total clinical effectiveness, alleviation of angina symptoms, and improvement in ECG performance.Conclusion:HBO combined with conventional drugs can better alleviate the symptoms of angina pectoris due to coronary heart disease and improve their ECG performance. In the future, high-quality RCTs need to be carried out to verify the conclusion of this study.

Objective:To systematically evaluate the efficacy of hyperbaric oxygen (HBO) combined with comentional drugs in the treatment of angina pectoris caused by coronary heart disease with meta-analysis.Methods:The data on clinical randomized controlled trials (RCTs) of HBO combined with conventional drugs in the treatment of angina pectoris due to coronary heart disease in Cochrane Library, Medline, Embase, Wanfang Data, Cqvip, China National Knowledge Infrastructure (CNKI) were collected, and the time period was from the respective founding day of the above databases to October 2020. Three researchers independently screened the collected RCTs, extracted relevant data, and evaluated the quality of the trials. RevMan 5.4 software was used for meta-analysis.Results:A total of 11 RCTs were included in this study. Meta-analysis results showed that the HBO group was better than the control group in terms of total clinical effectiveness, alleviation of angina symptoms, and improvement in ECG performance.Conclusion:HBO combined with conventional drugs can better alleviate the symptoms of angina pectoris due to coronary heart disease and improve their ECG performance. In the future, high-quality RCTs need to be carried out to verify the conclusion of this study.

Objective To explore the effect of mitral valve morphology on short-term and long-term outcomes in elderly patients with rheumatic mitral stenosis(MS)undergoing percutaneous balloon mitral valvuloplasty(PBMV).Methods In the prospective study,elderly patients with rheumatic mitral stenosis undergoing PBMV between February 1996 and June 2007 were followed up for 10 years.One hundred and twenty-four patients with full follow-up data were included in the study.According to echocardiography Wilkins score,83 cases of patients (Wilkins score ≤ 8)were selected as Wilkins score ≤ 8 group,and 41 cases were selected as Wilkins score＞ 8 group.The pressure gradient in mitral valve(MV△P),mitral valve area(MVA),left atrial diameter(LAD),left ventricular end-systolic diameter(LVESd),left ventricular end-diastolic diameter(LVEDd),pulmonary artery systolic blood pressure(sPAP),ejection fraction(EF)and mitral regurgitation were measured before and after the operation.Results In both Wilkins score ≤ 8 group and Wilkins score＞ 8 groups,MVA and EF were increased immediately after PBMV operation versus before PBMV operation,and MV△P,LAD,LVEDd,LVEDs and PAPs were decreased immediately after PBMV operation versus before PBMV operation(all P ＜0.05).Severe mitral regurgitation was not found in both two groups.The clinical effects of the Wilkins score≤8 group were stable after 10 years,which had no significant difference in the indexes compared with those in the Wilkins score≤ 8 group immediately after PBMV(P＞0.05).MVA and EF in the Wilkins score＞8 group were decreased,and MV△P,LAD,LVEDd,LVEDs and PAPS were increased after 10 years as compared with those immediately after PBMV(P＜0.05).The incidence of NYHA functional class Ⅲ or Ⅳ was lower in the Wilkins score ≤8 group than in the Wilkins score＞8 group(26.5％ or 22/83 vs.46.3％ or 19/41,x2 =4.879,P=0.027).And the incidence of mitral restenosis was lower in the ≤8 group than in the Wilkins score＞8 group(34.9％ or 29/83 vs.61.0％ or 25/41,x2 =7.567,P=0.006).There was no significant difference in the incidence of moderate or severe mitral regurgitation between the two groups(10.8％ or 9/83 vs.12.3％ or 5/41,x2=1.278,P=0.258).Conclusions The short-term and long-term outcomes are good in elderly individuals with rheumatic mitral stenosis undergoing PBMV operation,and the curative effect of PBMV operation is better in patients with Wilkins score ≤8 than in patients with Wilkins score ＞8.

Objective: To explore the clinical and molecular genetic features of a Chinese patient with catecholaminergic polymorphic ventricular tachycardia (CPVT). Methods: Clinical data including resting electrocardiography, echocardiography and treadmill exercise testing of a patient with CPVT admitted to our department in March 2013 were analyzed, and the peripheral venous blood samples of the patient and his family members and 400 ethnicity-matched healthy controls were obtained. All exons and exon-intron boundaries of the six CPVT-related genes including RYR2, CASQ2, TRDN, CALM1, KCNJ2 and ANKB were sequenced to detect the variants related to CPVT. The relationship between the genotypes and phenotypes was analyzed to direct the target therapy. Results: Recurrent syncope induced either by exercise or extreme frightened fear was observed in this patient. There was no positive family history of syncope or sudden death. The resting electrocardiography and echocardiography of the patient were normal, while the exercise testing revealed bidirectional and polymorphic ventricular tachycardia. A cardiac ryanodine receptor gene mutation (R2401H) was identified in this patient, while this mutation was absent in his parents and sister and 400 controls. No variant was detected in the remaining five candidate genes. Treatment with high dose of metoprolol succinate (118.75 mg/d) was effective and patient was free of syncopal attack during the 2 years follow-up. Conclusion This is the first report on RyR2-R2401H mutation in Chinese patient with CPVT, and high dose of metoptolol is the effective therapy option for CPVT related to RyR2 mutation.